番茄细菌性叶斑病菌的拮抗菌筛选、鉴定及其拮抗性能评价

doi:10.11686/cyxb2014450

摘要/Abstract

摘要： 采用含菌平板抑菌圈测定法,从120株高寒草地牧草内生细菌中分离筛选到一株对番茄细菌性叶斑病拮抗能力较强的菌株264ZY7,其抑菌圈直径为1.22 cm。264ZY7菌体短杆状,大小为1.534 μm×0.571 μm~3.210 μm×0.781 μm,革兰氏阳性菌,根据形态特征,并结合16S rDNA和gyrB基因序列相似性分析,将菌株264ZY7鉴定为解淀粉芽孢杆菌Bacillus amyloliquefaciens。通过室内盆栽防效试验,结果表明该菌株对番茄细菌性叶斑病的防效达69%,此外,该菌株对8种病原真菌具有抑菌能力,说明抑菌谱较宽,且抑菌率均大于30%,对番瓜根腐病菌(Fusarium sp.)效果最明显,抑菌率为52.84%。该研究结果为高寒草地紫花针茅内生细菌264ZY47的进一步开发利用提供了依据。

Abstract: After screening by assessing inhibition zones on nutrient media an antagonistic endophytic bacteria, strain 264ZY7, was identified as having strong antagonistic ability against Pseudomonas syringe pv. tomato which causes bacterial spot on tomatoes. This strain was selected from 120 endophytic bacteria strains isolated from alpine meadow pasture. Strain 264ZY7 produced a 1.22 cm diameter inhibition zone. The bacteria is rod-shaped with a spore size of 1.534 μm×0.571 μm-3.210 μm×0.781 μm, gram-positive and was identified as Bacillus amyloliquefaciens based on morphology and homology analysis of the 16S rDNA and gyrB gene sequence. The control efficacy of strain 264ZY7 on bacterial spot on tomatoes grown in pots ranged up to 69%. It was also able to control fungal pathogens, in particular Fusarium sp. (52.84% efficacy). These results suggest that further development and utilization of this endophytic bacteria strain (264ZY7) should be undertaken.

冯中红, 王玉琴, 杨成德, 薛莉, 陈秀蓉. 番茄细菌性叶斑病菌的拮抗菌筛选、鉴定及其拮抗性能评价[J]. 草业学报, 2015, 24(8): 166-173.

FENG Zhong-Hong, WANG Yu-Qin, YANG Cheng-De*, XUE Li, CHEN Xiu-Rong. Screening, identification and assessment of endophytic bacteria antagonistic totomato bacterial spot[J]. Acta Prataculturae Sinica, 2015, 24(8): 166-173.

参考文献

[1] Zhao T C, Yu L, Sun F Z, et al . Tomato bacterial speck disease and its prevention. Plant Protection, 1999, 25(4): 56.
[2] Yunis G P, Crall J M. First report of a bacreial speck of tomato by Pseudomonas syringa pv. tomato in Georgia. Plant Disease, 1980, 83: 199.
[3] Bonn, Martin H L. Bacterial spot of tomato leaves occurrence law in Frolidia. Journal of Experimental Agriculture, 1980, 39: 479-485.
[4] Zhao T C, Sun F Z, Song W S. Pathogen identification of bacterial speck of tomato. Acta Phytopathologica Sinica, 2001, 31(1): 37-42.
[5] Deng G, Qu X, Chen X R, et al . Pathogen identification of bacterial leaf speck of tomato based on 16S rDNA, physiology and biochemistry in Gansu. Plant Protection, 2008, 34(5): 47-51.
[6] Yi Y J, Luo K, Liu E M. Effect of endophytic bacterial on biological control of plant disease. Journal of Nuclear Agricultural Sciences, 2007, 21(5): 474-477.
[7] Li C S. A review on research progress of the biologicial control of soil-borne plant diseases with bacteria. Chinese Journal of Biological Control, 1992, 8(4): 168-172.
[8] Luo M Y, Lu X J, Liao X L. Research advances on application of endophytic bacteria. Modern Agricultural Science and Technology, 2010, (7): 11-12.
[9] Huang X, Xu L L, Huang R S, et al . Research advance in controlling plant diseases by Bacillus subtilis . Biotechnology Bulletin, 2010, (1): 24-28.
[10] Fang Z D. Research Method of Plant Pathology (Third Edition)[M]. Beijing: Chinese Agriculture Press, 1998: 110, 182-183.
[11] Gao X X, Man B Y, Chen X R, et al . Identification and determination of biological characteristics of Kobresia capillifolia endophytic bacteria X4 in the East Qilian Mountain Alpine grasslands. Acta Prataculturae Sinica, 2013, 22(4): 137-146.
[12] Zhao J, Lan C Z, Chen Q H, et al . Isolation of endophytic bacteria from tomato and selection of antagonistic bacteria to tomato bacterial wilt disease. Wuyi Science Journal, 2006, 22(1): 49-54.
[13] Li Z D, Chen X R, Yang C D. Identification of Polygonum viviparum endophytic bacteria Z17 and its capacity to antagonistic towards pathogenic fungi. Pratacultural Science, 2011, 28(12): 2096-2101.
[14] Wang L, Lee F, Tai C, et al . Comparison of gyrB gene sequences,16S rRNA gene sequences and DNA-DNA hybridization in the Bacillus subtilis group. International Journal of Systematic and Evolutionary Microbiology, 2007, 57: 1846-1850.
[15] Yamamoto S, Harayama S. PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Applied Environmental Microbiology, 1995, 61: 1104-1109.
[16] He H, Cai X Q, Hong Y C, et al . Selection of endophytic antifungal bacteria from capsicum. Chinese Journal of Biological Control, 2002, 18(4): 171-175.
[17] Li Q Q, Luo K, Lin W, et al . Isolation of tomato endophytic antagonists against Ralstonia solanacearum . Acta Phytopathologica Sinica, 2003, 33(4): 364-367.
[18] Huang M Y, Gu W J, Zhang F B, et al . Identification and fermentation of antagonistic bacterium against Ralstonia solanacearum . Microbiology, 2011, 38(2): 214-220.
[19] Li W J, Jiang R B. Isolation of tomato antagonists against Ralstonia solanacearum . Journal of Microbiology, 2007, 27(1): 5-8.
[20] Yang M L, Sun X L. Research of endophytic bacteria. Microbiology, 1998, 25(4): 224-227.
[21] Zhang J, Ran X X, Zhu T H, et al . Bacillus amyloliquefaciens antimicrobial effect on Pestalotiopsis guepini (Desm.) Stey. Journal of Northeast Forestry University, 2014, (7): 122-125.
[22] Chen S Y, Yang B Y, Gao M Y, et al . Inhibition of sclerotia formation of sclerotinia sclerotiorum by Bacillus amyloliquefaciens . Chinese Journal of Applied & Environmental Biology, 2005, 11(3): 373-376.
[23] Quan C S, Wang J H, Xu H T, et al . Identification and characterization of a Bacillus amyloliquefaciens with high antifungal activity. Acta Microbiologica Sinica, 2006, 46(1): 7-12.
[24] Zhao H, Lu C T, Liu H Y, et al . Identification of a Bacillus amyloliquefaciens strain and preliminary research on its antifungal activity. Journal of Henan Agricultural Sciences, 2014, 43(4): 75-79.
[25] Zhao X Y, Yang H, Chen J G, et al . Screening and fermentation optimization of antagonistic bacillus against tobacco bacterial wilt. Hubei Agricultural Sciences, 2014, (8): 1810-1814.
[26] Fan X J, Yang C Q, Qiu S X, et al . Identification of biocontrol bacterial strain and its antagonistic activity to tomato bacterial speck and colonizing ability in tomato. Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2013, 42(4): 337-341.
[1] 赵廷昌, 于莉, 孙福在, 等. 番茄细菌性斑点病及其防治. 植物保护, 1999, 25(4): 56.
[4] 赵廷昌, 孙福在, 宋文生. 番茄细菌性斑点病病原菌鉴定. 植物病理学报, 2001, 31(1): 37-42.
[5] 邓刚, 屈星, 陈秀蓉, 等. 甘肃番茄细菌性斑点病病原菌鉴定. 植物保护, 2008, 34(5): 47-51.
[6] 易有金, 罗宽, 刘二明. 内生细菌在植物病害生物防治中的作用. 核农学报, 2007, 21(5): 474-477.
[7] 李长松. 拮抗性细菌生物防治植物土传病害的研究进展. 生物防治通报, 1992, 8(4): 168-172.
[8] 罗茗月, 路雪君, 廖晓兰. 植物内生细菌的应用研究进展. 现代农业科技, 2010, (7): 11-12.
[9] 黄曦, 许兰兰, 黄荣韶,等. 枯草芽孢杆菌在抑制植物病原菌中的研究进展. 生物技术通报, 2010, (1): 24-28.
[10] 方中达. 植病研究方法(第三版)[M]. 北京: 中国农业出版社, 1998: 110, 182-183.
[11] 高晓星, 满百膺, 陈秀蓉, 等. 东祁连山线叶嵩草内生细菌X4的产吲哚乙酸、解磷、抗菌和耐盐特性研究及分子鉴定. 草业学报, 2013, 22(4): 137-146.
[12] 赵健, 兰成忠, 陈庆河, 等. 抗番茄青枯病内生拮抗细菌的筛选及室内防效测定. 武夷科学, 2006, 22(1): 49-54.
[13] 李振东, 陈秀蓉, 杨成德. 珠芽蓼内生菌Z17抑菌能力测定及其鉴定. 草业科学, 2011, 28(12): 2096-2101.
[16] 何红, 蔡学清, 洪永聪, 等. 辣椒内生细菌的分离及拮抗菌的筛选. 中国生物防治, 2002, 18(4): 171-175.
[17] 黎起秦, 罗宽, 林纬, 等. 番茄青枯病内生拮抗细菌的筛选. 植物病理学报, 2003, 33(4): 364-367.
[18] 黄明媛, 顾文杰, 张发宝, 等. 番茄青枯病拮抗菌筛选鉴定及其发酵条件初探. 微生物学通报, 2011, 38(2): 214-220.
[19] 李伟杰, 姜瑞波. 番茄青枯病拮抗菌的筛选. 微生物学杂志, 2007, 27(1): 5-8.
[20] 杨梅莲, 孙晓璐. 植物内生细菌的研究. 微生物学通报, 1998, 25(4): 224-227.
[21] 张静, 冉晓潇, 朱天辉, 等. 解淀粉芽孢杆菌对山茶灰斑病菌的抑制作用. 东北林业大学学报, 2014, (7): 122-125.
[22] 陈士云, 杨宝玉, 高梅影, 等. 一株抑制油菜核盘菌菌核形成的解淀粉芽孢杆菌. 应用与环境生物学报, 2005, 11(3): 373-376.
[23] 权春善, 王军华, 徐洪涛, 等. 一株抗真菌解淀粉芽孢杆菌的分离鉴定及其发酵条件的初步研究. 微生物学报, 2006, 46(1): 7-12.
[24] 赵辉, 鲁传涛, 刘红彦, 等. 一株生防芽孢杆菌的鉴定及其抑菌活性的初步研究. 河南农业科学, 2014, 43(4): 75-79.
[25] 赵晓燕, 杨欢, 陈建刚, 等. 一株烟草青枯病生防菌的筛选鉴定及其发酵优化. 湖北农业科学, 2014, (8): 1810-1814.
[26] 范晓静, 杨春泉, 邱思鑫, 等.番茄细菌性斑点病生防菌的鉴定、防病及定殖力. 福建农林大学学报(自然科学版), 2013, 42(4): 337-341.
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